Method for quantifying human reticulocytes

Abstract

A method for determining the reticulocyte population in a blood sample includes the steps of determining the average cell size of the blood sample, partitioning the sample by centrifugation through a medium of known density so as to provide a fraction enriched with neocytes, determining the average cell size of the fraction, comparing the average cell size of the sample to the average cell size of the fraction and utilizing this comparison to provide a determination of the population of reticulocytes in the sample.

We claim: 
     
       1. A method for determining the reticulocyte population in a blood sample of a patient not having a hematological disease characterized by an abnormally dense red cell population, comprising the steps of: (a) determining the average cell size of the blood sample;   (b) partitioning the sample by centrifugation through a medium of known density equal to a specific density of 1.089 so as to provide a fraction enriched with neocytes;   (c) determining the average cell size of the fraction obtained in step (b); and   (d) comparing the average cell size of the sample, determined in step (a), to the average cell size of the fraction, determined in step (c), and utilizing this comparison to provide a determination of the population of reticulocytes in the sample by correlating the comparison to a set of predetermined measurements of reticulocyte population and corresponding comparison values.   
     
     
       2. The method of claim 1 in which the medium of known density permits concentration of neocytes therein during centrifugation. 
     
     
       3. The method of claim 1 in which the medium of known density is arabino-galactan polysaccharide. 
     
     
       4. The method of claim 1 in which the determination of average cell size is made utilizing a blood cell analyzer to provide a measurement of mean corpuscular volume. 
     
     
       5. A method for quantifying the reticulocyte population in a blood sample of a patient not having a hematological disease characterized by an abnormally dense red cell population, comprising the steps of: (a) determining the means corpuscular volume of the sample;   (b) subjecting the sample to density medium fractionation utilizing a medium having a specific density of 1.089 to yield a fraction which is enriched with neocytes;   (c) determining the mean corpuscular volume of the fraction of step (b);   (d) comparing the mean corpuscular volume of the fraction, determined in step (c), to the mean corpuscular volume of the sample, determined in step (a), and utilizing this comparison to provide an indication of the reticulocyte population in the sample by correlating the comparison to a set of predetermined measurements of reticulocyte population and corresponding comparison values.   
     
     
       6. The method of claim 5 in which the density medium fractionation step utilizes a medium which permits concentration of neocytes therein during centrifugation. 
     
     
       7. The method of claim 5 in which the density medium fractionation step utilizes arabino-galactan polysaccharide. 
     
     
       8. A method for determining the reticulocyte population in a blood sample of a patient not having a hematological disease characterized by an abnormally dense red cell population, comprising the steps of: (a) diluting the sample;   (b) determining the mean corpuscular volume of the sample;   (c) centrifuging the sample with a medium of known density equal to a specific density of 1.089 to provide a fraction enriched with neocytes;   (d) determining the mean corpuscular volume of the fraction of step (c);   (e) comparing the mean corpuscular volume of the sample to the mean corpuscular volume of the fraction and correlating this comparison to a standard set of measurements of reticulocyte population and corresponding mean corpuscular volume comparison values to achieve a determination of the reticulocyte population of the sample.   
     
     
       9. The method of claim 8 in which the medium of known density permits concentration of neocytes therein during centrifugation. 
     
     
       10. The method of claim 8 in which the medium of known density is arabino-galactan polysaccharide. 
     
     
       11. The method of claim 8 in which the determination of mean corpuscular volume is made utilizing a blood cell analyzer. 
     
     
       12. A method for determining the reticulocyte population in a blood sample of a patient not having a hematological disease characterized by an abnormally dense red cell population, comprising the steps of: (a) determining the average cell size of the blood sample;   (b) determining the average cell size of a fraction of the sample having a specific density of 1.089; and   (c) comparing the average cell size of the sample, determined in step (a), to the average cell size of the fraction, determined in step (b), and utilizing this comparison to provide a determination of the population of reticulocytes in the sample by correlating the comparison to a set of predetermined measurements of reticulocyte population and corresponding comparison values.   
     
     
       13. The method of claim 12 including the step of determining average cell size by utilizing a blood cell analyzer to provide a measurement of mean corpuscular volume. 
     
     
       14. The method of claim 12 including the step of determining the average cell size by the mean corpuscular volume. 
     
     
       15. The method of claim 12 including the step of diluting the sample prior to step (a). 
     
     
       16. The method of claim 12 including the step of partitioning the sample by centrifugation through a medium with a specific density of 1.089 to provide the fraction.